The functional properties of the CD172a− CD24+ cells within the CD4−8− cDC subset preparations. The cDC subsets were isolated as in Figure 1 using the NK-marker depletion protocol, then the CD172a positive and negative fractions of the CD4−8−CD11chi subset separated as in Figure 4. (A) Production of IL-12p70 following stimulation with CpG 1668, IL-4, GM-CSF, and IFN-γ. Production is from 105 cells in 0.2 mL medium for 40 hours. The mean plus or minus SD of 3 cultures is given for one experiment, typical of 2. (B) Ability to process and cross-present on MHC I soluble antigen in vitro, assayed by culture of freshly isolated cDC fractions with ovalbumin and OT-I CD8 T cells. Points are the mean of 4 cultures plus or minus SD. One experiment typical of 2 is shown. (C) Ability to process and cross-present soluble antigen on MHC I in vivo, assayed by isolation of cDC fractions 20 hours after injection of mice with lipopolysacchride followed by ovalbumin, then culture of the cDC fractions with OT-I CD8 T cells without addition of ovalbumin. The result is a single experiment. Points are the mean of 3 to 4 cultures plus or minus SD.